Octocorallia: Gorgoniidae) from Ecuador, with a Description of a New Species

Scientia Marina 80(3) September 2016, 000-000, Barcelona (Spain) ISSN-L: 0214-8358 doi: http://dx.doi.org/10.3989/scimar.04392.14A

New records of the genera Leptogorgia, Pacifigorgia and Eugorgia (Octocorallia: Gorgoniidae) from Ecuador, with a description of a new species

María del Mar Soler-Hurtado 1,2, Annie Machordom 1, Jesús Muñoz 3,4, Pablo J. López-González 2 1 Dpto. Biodiversidad y Biología Evolutiva. Museo Nacional de Ciencias Naturales (MNCN-CSIC), 28006 Madrid, Spain. E-mail: [email protected]; [email protected] 2 Biodiversidad y Ecología de Invertebrados Marinos, Facultad de Biología, Universidad de Sevilla, 41012, Sevilla, Spain. E-mail: [email protected] 3 Real Jardín Botánico (CSIC), Plaza de Murillo 2, 28014 Madrid, Spain. E-mail: [email protected] 4 Centro de Biodiversidad y Cambio Climático, Universidad Tecnológica Indoamérica, Quito, Ecuador.

Summary: New records of the genera Leptogorgia, Pacifigorgia and Eugorgia (Octocorallia: Gorgoniidae) on the coast of Ecuador are reported. These new records redefine the current known limit of distribution of these species on the eastern Pacific coast (from southern California to Chile). Some of these species are reported for the first time since their original description. The newly collected specimens allow for the measurement of the variability of several morphological characters, from colonial to sclerite levels. Additionally, Pacifigorgia machalilla n. sp. is described and compared with its closest rela- tives. Morphological differentiation among related species is supported by genetic divergence estimated from an extended barcode of MutS + Igr + COI. Keywords: Gorgoniidae; eastern Pacific; distribution; mtMutS; COI; barcode; Igr. Nuevos registros de los géneros Leptogorgia, Pacifigorgia y Eugorgia (Octocorallia: Gorgoniidae) en Ecuador, con la descripción de una especie nueva Resumen: Se han obtenido nuevos registros para los géneros Leptogorgia, Pacifigorgia y Eugorgia (Octocorallia: Gorgo- niidae) en la costa de Ecuador. Estos nuevos registros redefinen el límite actual conocido en la distribución de estas especies en la costa del Pacífico Oriental (desde el sur de California hasta Chile). Algunas de estas especies han sido registradas por primera vez desde su descripción original. Las nuevas muestras recogidas permiten medir la variabilidad de los caracteres morfológicos, desde el nivel de colonia hasta el de esclerito. Además, se describe la especie Pacifigorgia machalilla n. sp., y se compara con sus congéneres más relacionados. Las diferenciaciones morfológicas encontradas entre estas especies, están apoyadas por la divergencia genética estimada a partir del “código de barras” ampliado, formado por los genes MutS + Igr + COI. Palabras clave: Gorgoniidae; este del Pacífico; distribución; mtMutS; COI; “código de barras”; Igr. Citation/Como citar este artículo: Soler-Hurtado M.M., Machordom A., Muñoz J., López-González P.J. 2016. New records of the genera Leptogorgia, Pacifigorgia and Eugorgia (Octocorallia: Gorgoniidae) from Ecuador, with a description of a new species. Sci. Mar. 80(3): 000-000. doi: http://dx.doi.org/10.3989/scimar.04392.14A LSID: urn:lsid:zoobank.org:pub:A6373513-5FFE-4D2E-B130-98C239C0EDF3 Editor: X. Turon. Received: December 18, 2015. Accepted: June 1, 2016. Published:September 25, 2016. Copyright: © 2016 CSIC. This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-by) Spain 3.0 License.

INTRODUCTION gorgonian octocorals are one of the best-represented taxonomic groups in sublittoral marine ecosystems, The octocorals are found in marine habitats rang- are ecologically important and are the dominant mac- ing from intertidal to abyssal waters and are distributed rofaunal group on many tropical reefs (Sánchez et al. from the Arctic to the Antarctic (Bayer 1961). The 2003, Williams and Breedy 2004). The study of the 2 • M.M. Soler-Hurtado et al. eastern Pacific gorgonian octocorals, specifically in Ecuador, has not matched the intensity and number of publications dedicated to Caribbean species (see Bayer 1981). Despite important contributions by authors such as Breedy and Guzmán (2002, 2007), Williams and Breedy (2004) and Breedy et al. (2009) in the eastern Pacific, the knowledge on the gorgonian fauna of Ecuador is far from complete (but see Bielschowsky 1929 and Soler-Hurtado and López-González 2012). Four gorgoniid genera (Gorgoniidae) were previously reported in the eastern Pacific, Phycogorgia Milne-Ed- wards and Haime, 1850, Leptogorgia Milne-Edwards and Haime, 1857, Eugorgia Verrill, 1868 and Paci figorgia Bayer, 1951. The genus Leptogorgia, with about 60 valid species (27 species in the eastern Pa- cific) is one of the most frequent genera in the shallow Fig. 1. – Station map in Ecuadorian waters where gorgonians octoc- water communities of the eastern Pacific (Breedy and orals were sampled from February 2010 to June 2014. Guzmán 2007), with a wide distribution from southern California to Chile. This genus is also present in the took colour photographs of the living specimens in Caribbean, western and eastern South African coasts, their environment; specimens were also photographed the eastern Atlantic and the Mediterranean Sea, and one outside the water, just after collection to ensure the species is known from the sub-Antarctic (Williams and accuracy of our colour descriptions. Subsamples were Lindo 1997). The genus Pacifigorgia is geographically fixed in either absolute ethanol for further molecular confined to the Pacific coast of tropical America, with analyses, or in 4% buffered formalin (after previous the exception of Pacifigorgia elegans (Milne-Edwards relaxation with menthol crystals for a few hours) for and Haime, 1857) from the tropical western Atlantic. the morphological study; the rest of the colonies were Pacifigorgia includes about 36 recognized species dis- allowed to air dry. Buffered formalin-fixed subsamples tributed throughout the eastern tropical Pacific region were subsequently transferred to 70% ethanol (Soler- (Breedy and Guzmán 2002, 2003, 2004, Williams and Hurtado and López-González 2012). Breedy 2004, Guzmán and Breedy 2011). The genus Eugorgia, which includes 16 species, is considered ex- External morphology and SEM study clusive to the eastern Pacific (from southern California to Peru) (Breedy and Guzmán 2013). Finally, the ge- Fragments from different parts of the colony were nus Phycogorgia, with only one species, Phycogorgia prepared for study by SEM according to standard fucata (Valenciennes, 1846), is distributed along the methods (Bayer and Stefani 1989, Alderslade 1998). west coast of Central and South America (Bayer 1953). Additionally, permanent mounts were made for light The aim of this report is to present new informa- microscopy observation. The colonies are described tion derived from material of the genera Eugorgia, and illustrated according to standard terminology Leptogorgia and Pacifigorgia collected in Ecuador. (Bayer et al. 1983, Breedy and Guzmán 2003, 2007). We expand the known distribution of some species in For either morphological and molecular compari- the eastern Pacific and describe new morphological sons, we studied types and additional materials depos- variability for several taxa. Finally, using morphologi- ited in the Museum of Comparative Zoology, Harvard cal and genetic data, we describe one new species of University (MCZ); Muséum National d’Histoire Na- Pacifigorgia, emphasizing the set of morphological turelle, Paris (MNHN); the Natural History Museum, characters considered to be of taxonomic importance London (NHM); the Yale Peabody Museum of Natural for this genus (Breedy and Guzmán 2002, 2003, 2004, History, New Haven (YPM); and the National Museum Williams and Breedy 2004, Guzmán and Breedy of Natural History, Smithsonian, Washington (MNH). 2011). We include the sequence analysis of the barcode Museum specimens were revised for morphological of mtMutS plus COI with a short, adjacent intergenic comparative purposes and, in some cases, for molecular region (Igr1) proposed by McFadden et al. (2011), analysis. These samples are listed in the text after the with its closest congeners and available sequences in newly collected material of each species. Additional GenBank. museum specimens of species revised but not found in Ecuador were the following: MATERIALS AND METHODS Eugorgia ampla (Verrill, 1864): NHM 69.4.15.53, Baja California (Mexico), no further data. YPM 399, Study area and sampling methodology Baja California, 11-15 m depth, 1865. MCZ 65167, Mexico, no depth given, date unknown. Eleven stations in Ecuadorian waters were sampled Eugorgia aurantica (Horn, 1860): YPM (4051), from February 2010 to June 2014 (Fig. 1). Gorgonians Baja California (Mexico), no depth given, 1867-1870. were collected by SCUBA diving in a depth range of NHM 40.8.22.8, Baja California (Mexico), no depth between 5 and 30 m. Substrata in these habitats mainly given, date unknown. MCZ 36185, Baja California include sand and rocky bottoms. During sampling, we (Mexico), no depth given, date unknown.

SCI. MAR. 80(3), September 2016, 000-000. ISSN-L 0214-8358 doi: http://dx.doi.org/10.3989/scimar.04392.14A Gorgoniidae from Ecuador • 3

Table 1. – Pacifigorgia species involved in the molecular comparisons carried out in this paper. Materials in bold are species sequenced for this study. Note that all GenBank sequences are considered here with the names as they appear in GenBank and their original publications (including numbers or letters). For sequences with duplicate complete names, we have included (1), (2), (3), etc., for the purpose of correctly identifying the sequence in Table 5. Species Catalog. Nos Igr + COI mtMutS Pacifigorgia bayeri voucher HGM77 HG917061 HG917044 Pacifigorgia catedralensis (1) voucher HGM109 HG917065 HG917019 Pacifigorgia catedralensis (2) voucher HMG112 HG917066 HG917020 Pacifigorgia exilis YPM 4059 KX351878 KX351871 Pacifigorgia firma MCZ 51918 KX351879 KX351872 Pacifigorgia irene voucher HMG10 HG917070 HG917024 Pacifigorgia irene MNCN 2.04/1174 KX351880 KX351873 Pacifigorgia machalilla n. sp. (1) MECN Ant0058 KX351881 KX351874 Pacifigorgia machalilla n. sp. (2) MECN Ant0059 KX351882 KX351875 Pacifigorgia machalilla n. sp. (3) MECN Ant0061 KX351883 KX351876 Pacifigorgia media Parrin et al. 2009 GQ342421 GQ342497 Pacifigorgia sculpta MCZ 57053 KX351884 KX351877 Pacifigorgia smithsoniana voucher HMG59 HG917076 HG917023 Pacifigorgia stenobrochis voucher HMG100 HG917078 HG917026 Pacifigrogia rubicunda (1) voucher HMG01 HG917073 HG917032 Pacifigrogia rubicunda (2) voucher HMG29 HG917074 HG917033

Eugorgia nobilis Verrill, 1868: YPM 1552a-e, The newly collected specimens are deposited in the type material, Las Perlas (Panama), 11-15 m depth, Museo Ecuatoriano de Ciencias Naturales (MECN), 1866.YPM 401, type material, Baja California (Mex- the octocoral reference collection of the research group ico), no further data. MCZ 36316, Baja California “Biodiversidad y Ecología de Invertebrados Marinos” (Mexico), no further data. USNM 49371, Gulf of at the University of Seville (BEIM), the Museo Nacion- Nicoya (Costa Rica), no depth given, 15 Jan. 1930. al de Ciencias Naturales in Madrid (MNCN-CSIC), USNM 49372, Baja California (Mexico), no depth the Museo de Zoología de la Universidad Tecnológica given, 16 Mar. 1911. Indoamérica in Quito (UTI), and the Museu de Ciénces Eugorgia multifida Verrill, 1870: YPM 4605, type Naturals in Barcelona (BCN). material, Mazatlan (Mexico), no depth given, date unknown. USNM 49368, Baja California (Mexico), 11- DNA extraction, PCR amplification and 15 m depth, date unknown. sequencing Pacifigorgia adamsii (Verrill, 1868): MCZ 391, type material, Panama, no depth given, 1863. MCZ DNA was extracted from 20-30 mg of tissue using 36079, type material, Panama, no depth given, 1866- the DNeasy extraction kit (Qiagen, Inc.) according to 1867. NHM 1930.6.173.9, South Pacific, no depth giv- the manufacturer’s protocol. Partial COII-COI (includ- en, date unknown. YPM 1173g, type material, Panama, ing Igr1 region) and MutS sequences were amplified no depth given, 1866-1867. USNM 49688, Las Perlas by PCR using the following primers: COII8068F (Mc- (Panama), 33 m depth, 5 Mar 1888. USNM 49586, Fadden et al. 2004), COIOCTR (France and Hoover Mexico, no depth given, date unknown. 2002), the newly developed COI-Gorg1-R3 (5′-AGA- Pacifigorgia agassizii (Verrill, 1864): MCZ 374, GAAGGTGGTAATAACCAGAAA-3′) and COI- type material, Panama, no depth given, 1863. MCZ Gorg2-F2 (5′-GATTCGGAAATTGGTTTGTG-3´) 4035, Acapulco (Mexico), no depth given, 1859-1860. for COI + Igr1; ND42599F (France and Hoover 2002) MCZ 36266, Baja California (Mexico), no depth giv- and MUT3458R (Sánchez et al. 2003) for MutS. Am- en, date unknown. NHM 1885.5.18.2, type material, plifications were carried out in a final volume of 50 µL Mexico, no depth given, 1859-1860. YPM 956, type containing 5 µL of 10x buffer (containing 10×2 mM material, Baja California (Mexico), no depth given, MgCl2), 1 µL dNTPs mix (10 mM), 0.8 µL of each 1860. MNHN-IK 1589, no depth given, date unknown. primer (10 µM), 0.5 µL of Taq DNA polymerase (5U/ USNM 49369, Baja California (Mexico), no depth µL) (Biotools) and 2 µL of genomic DNA. Thermocy- given, date unknown. cling for the COI fragment included an initial 4-min Pacifigorgia arenata (Valenciennes, 1846): denaturation step at 94°C, followed by 40 cycles of 45 MNHN-IK 1411, type material, New Zealand, no s at 94°C, 1 min at 58°C and 1 min at 72°C. The cy- depth given, 1839. cle ended with 10 min of sequence extension at 72°C. Pacifigorgia firma Breedy and Guzmán, 2003: For MutS, we used an initial 4-min denaturation step MCZ 51918, type material, Culebra Bay (Costa Rica), at 94°C, followed by 35 cycles of 90 s at 94°C, 90 s 20 m depth, 21 Sept. 1997. at 58°C and 1 min at 72°C. The cycle ended with 5 Pacifigorgia pulchra var. exilis (Verrill, 1870): min of sequence extension at 72°C. The amplification YPM 4059, type material, Baja California (Mexico), products were purified by ethanol precipitation. The no depth given, 1867-1870. USNM 75076, Baja Cali- amplicons were sequenced for both strands using Big- fornia (Mexico), no depth given, date unknown. Dye Terminator in an ABI 3730 genetic analyser (Ap- Pacifigorgia rutila (Verrill, 1868): MCZ 184, type plied Biosystems). The sequences obtained were edited material, Acapulco (Mexico), no depth given, date un- using the Sequencher v.4.6 program (Gene Code Cor- known. YPM 2266, type material, Acapulco (Mexico), poration, Ann Arbor, MI, USA). The resulting align- no depth given, 1869-1870. ments were inspected by eye and manually checked

SCI. MAR. 80(3), September 2016, 000-000. ISSN-L 0214-8358 doi: http://dx.doi.org/10.3989/scimar.04392.14A 4 • M.M. Soler-Hurtado et al. and adjusted with Se-Al v2.0a11 (Rambaut 2002). The distance matrix was obtained using PAUP*v4.0b10 (Swofford 2001) with a neighbour joining clustering algorithm (Saitou and Nei 1987). A molecular data matrix was created with the morphologically closest species, together with other published sequences for Gorgoniidae in GenBank (see Table 1).

RESULTS

Order ALCYONACEA Verrill, 1866 Suborder Holaxonia Studer, 1887 Family Gorgoniidae Lamouroux, 1812

Genus Leptogorgia Milne-Edwards and Haime, 1857

Leptogorgia alba (Duchassaing and Michelotti, 1864) (Figs 2, 3)

Synonymy. See Breedy and Guzmán (2007: 12). Newly collected examined material: MECN (Ant0001), Los Ahor- cados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 15 m depth, 27 Feb. 2010, three whole colonies. MECN (Ant0019), Isla de Salango, Manabí (Ecuador), 1°35’55.13”S 80°52’0.01”W, 7 m depth, 20 Nov. 2011, two whole colonies. MECN (Ant0020), Salinas, Santa Elena (Ecuador), 2°12’50.01”S 80°56’5.93”W, 15 m depth, 18 Dec. 2011, one whole colony. MECN (Ant0027), Los Frailes, Manabí (Ecuador), 1°30’14”S 80°48’33”W, 10 m depth, Dec. 2011, three whole colonies. MECN (Ant0030), Los Frailes, Manabí (Ecuador), 1°30’14”S 80°48’33”W, 15 m depth, 19 March. 2012. BEIM (0071), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 15 m depth, 27 Feb. 2010, one whole colony. BEIM (0074), Los Ahor- cados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 5 m depth, 27 Feb. 2010, one whole colony. MNCN (2.04/482), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, Feb. 2010, one whole colony. MNCN (2.04/483), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 15 m depth, 28 Feb. 2010, one whole colony. MZB (2016-2989), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 15 m depth, 27 Feb. 2010, one whole colony. UTI (MZUTI-Inv02), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 15 m depth, 28 Feb. 2010, one whole colony. MECN (Ant0040), Punta Mala, Manabí (Ecuador), 1°33’41.37”S 80°50’8.79”W, 16 m depth, 02 Feb. 2013, one whole colony. MECN (Ant0041), Islote La Viuda, Manabí (Ecuador), 1°26’5.95”S 80°46’7.30”W, 15 m depth, 23 Nov. 2013, one whole colony. MECN (Ant0050) Punta Machalilla, Manabí (Ecuador), 1°28’33.53”S 80°47’38.04”W, 13 m depth, 24 Nov. 2013, one whole colony. Additional examined materials: Litigorgia laevis, MCZ 5416, type material, Las Perlas (Panama), no depth given, date unknown; MCZ 7008, type material, Golfo de Nicoya, (Costa Rica), no depth given, May 1868. Leptogorgia laevis, YPM IZ 001639, type material, Las Perlas, (Panama), no depth given, 1868. Leptogorgia alba, NHM 1946.1.14.52, Toboga (Panama), no depth given, date unknown; NHM 30.6.17.13, Toboga (Panama), no depth given, date unknown. Leptogorgia alba, USNM 59078, Ecuador, 8-9 m depth, 8 May 1966; USNM 1016223, Ecuador, no depth given, Dec. 1937.

Description. The colonies measure up to 350 mm in length and 105 mm wide, irregularly pinnate; branches slender, mostly in a plane (Fig. 2A). Unbranched distal twigs up to 100 mm in length and 20 mm in diameter, compressed proximally, more cylindrical and slightly tapered distally (Fig. 2A, B). The holdfast circular, up to 5 mm in diameter. Slightly marked longitudinal grooves along the thick basal branches and near the base. The polyps retract within slightly raised polyp- mounds, sparsely distributed all around the branches Fig. 2. – Leptogorgia alba (MNCN 2.04/482). A, colony; B, detail with oblong apertures (Fig. 2B). The colour of the of a branch; C, light micrograph of sclerites.

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Costa Rica, Panama, Colombia, and the Galápagos Islands (Ecuador) (Duchassaing and Michelotti 1864, Bielschowsky 1929, Breedy and Guzmán 2007), at 3-30 m depth (Fig. 25A). Although Leptogorgia alba has already been collected in Ecuador (Galápagos Islands) (Breedy and Guzmán 2007); this is the first time that it has been found on the continental coast since the paper of Biels- chowsky (1929).

Remarks. Our material is in agreement with the original description (Duchassaing and Michelotti 1864:19) and its later re-description (Breedy and Guzmán 2007:12-19). Due to its white colour, it is easily recognizable underwater. Breedy and Guzmán (2007) considered that its morphological variability, especially the type of branching, could perhaps be a response to several environmental factors. In our samples we observe a more pinnate pattern, although there is a tendency towards a dichotomous branching in some specimens. In the type material, the spindles are long, up to 0.17- 0.18 mm in length and 0.04-0.06 mm in width, with marked and complex tubercles. However, our material shows smaller spindles (up to 0.14×0.03 mm, Table 2) with a less marked ornamentation. In the same way, the anthocodial rods are considered very consistent in size and shape (long and conspicuous) in this species (up to 0.15 mm in length) (Breedy and Guzmán 2007). How- ever, the examined Ecuadorian material shows a smaller maximum length (up to 0.08 mm).

Leptogorgia diffusa (Verrill, 1868) (Figs 4, 5)

Synonymy. See Breedy and Guzmán (2007: 32). Newly collected examined material: MECN (Ant0032), Punta Mala, Manabí (Ecuador), 1°33’41.37”S 80°50’8.79”W, 13 m depth, 3 Abr. 2012, one whole colony. MECN (Ant0005), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 30 m depth, 28 Feb. 2010, two whole colonies. BEIM (0080), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, Feb. 2010, one whole colony. BEIM (0078), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 20 m depth, 28 Feb. 2010, one whole colony. MNCN (2.04/484), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 15 m depth, 27 March 2012, one whole colony. MNCN (2.04/485), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 30 m depth, 27 Feb. 2010, one whole colony. MZB (2016-2990), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 30 m depth, 27 Feb. 2010, one whole colony. MZB (2016-2991), Los Ahorcados, Manabí (Ecuador), Fig. 3. – Leptogorgia alba (MNCN 2.04/482) SEM photographs. 1°40’44”S 80°50’08”W, 25 m depth, 27 Feb. 2010, one whole Coenenchymal sclerites, A, spindles; B, captans; C, anthocodial colony. UTI (MZUTI-Inv09), Los Ahorcados, Manabí (Ecuador), sclerites, rods. 1°40’44”S 80°50’08”W, 30 m depth, 28 Feb. 2010, one whole colony. UTI (MZUTI-Inv01), Los Ahorcados, Manabí (Ecuador), colony is white. The coenenchymal sclerites hyaline 1°40’44”S 80°50’08”W, 10 m depth, 27 Feb. 2010, one whole (Fig. 2C). The dominant sclerite type spindles up to colony. MECN (Ant0042), El Burbullón, Manabí (Ecuador), 1°28’23.59”S 80°51’23.04”W, 25 m depth, 23 Nov. 2013, one 0.14 mm in length and 0.03 mm wide, with 4-6 whorls whole colony. of tubercles; straight or bent, some with a marked waist (Figs 2C, 3A). The capstans up to 0.07 mm in length Additional examined materials: Leptogorgia diffusa MCZ 7081, type material, Golfo de Nicoya (Costa Rica), no depth given, May and 0.03 mm wide (Fig. 3B). Crosses not found. The 1868. Litigorgia diffusa YPM 1659A, type material, Las Perlas anthocodial sclerites hyaline rods up to 0.08 mm in (Panama), no depth given, 1866-1867. Leptogorgia diffusa MCZ length and 0.02 mm wide, with some marginal projec- 4972, Mexico, no depth given, date unknown. MNHN-IK 2233, no tions (Figs 2C, 3C). further data.

Geographic and bathymetric distribution. Lepto Description. The colonies are up to 670 mm in gorgia alba has been reported in Mexico, El Salvador, length by 950 mm in width. The branching pattern

SCI. MAR. 80(3), September 2016, 000-000. ISSN-L 0214-8358 doi: http://dx.doi.org/10.3989/scimar.04392.14A 6 • M.M. Soler-Hurtado et al. yes yes yes pinnae 0.15×0.05 0.08×0.05 0.14×0.06 0.07×0.06 prominent red, yellow double disc dark orange not obtained Eugorgia daniana - - - no red purple spindles 0.13×0.04 0.08×0.04 0.06×0.04 prominent dichotomous orange / 0.07×0.02 Leptogorgia obscura - - - flat yes collected in this study, on the coast of Ecuador. brown capstans 0.1×0.03 0.07×0.03 0.03×0.03 pink / 0.16×0.08 irregularly pinnate Leptogorgia flexilis red, pink and yellow Eugorgia and Leptogorgia - - - no red red spindles 0.11×0.03 0.07×0.04 0.04×0.05 prominent flat, lax, pinnate orange / 0.13×0.02 Leptogorgia diffusa - - - no no white spindles colourless 0.14×0.03 0.07×0.03 slightly raised Leptogorgia alba irregularly pinnate colourless / 0.08×0.02 Comparative general features of the species genera Fig. 4. – Leptogorgia diffusa (MNCN 2.04/1172). A, colony; B, detail of a branch; C, light micrograph of sclerites.

is irregularly pinnate; branches are flat, lax, pinnate, Table 2. – slender and on a plane (Fig. 4A). The unbranched distal twigs can reach up to 22 mm in length and 17 mm in diameter (Fig. 4A, B). The holdfast is slightly flat, up to 15 mm in diameter. The polyps retract within prominent polyp-mounds, sparsely distributed all around the branches with slit-like apertures (Fig. 4B). The colour of the colony and of the coenenchymal sclerites is red (Fig. 4A-C). The spindles are the dominant sclerite Characters Colour of colony Type of branching Polyp-mounds Dominant sclerite type Colour of sclerites Bicolour sclerites Acute spindles max. size (mm) Capstan max. size (mm) Crosses (mm) Size and colour of anthocodial rods (mm) Complete double disc Double disc max. size (mm) Disc-spindles type, up to 0.11 mm in length and 0.03 mm in width,

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with 4-6 whorls of tubercles, they are straight or bent, some with a marked waist (Figs 4C, 5A). The capstans reach up to 0.07 mm in length and 0.04 mm in width (Figs 4C, 5B). Some small, scattered crosses are found (0.04×0.05 mm) (Fig. 4C). The anthocodial sclerites are orange flattened rods, up to 0.13 mm in length and 0.02 mm in width, with lobe-like marginal projections (Figs 4C, 5C).

Geographic and bathymetric distribution. Lepto gorgia diffusa has been previously reported in Califor- nia, El Salvador, Costa Rica, Panama, Colombia, and Chile at 5-30 m depth (see Verrill 1868, Bielschowsky 1929, Prahl et al. 1986, Breedy and Guzmán, 2007) (Fig. 25B). This new record fills the gap between the northern and southern records of the species. Lepto gorgia diffusa probably has a wider distribution, especially in offshore areas of Mexico and Peru, but it may have been previously overlooked. Although this species is quite frequent in Ecuador, it is usually observed isolated within multispecies assemblages and does not form large gorgonian gardens.

Remarks. The morphology of this species is very constant in all examined samples. It is easily differenti- able from other Leptogorgia species by the lax, pinnate style of branching, with prominent polyp-mounds. Breedy and Guzmán (2007: 34-35) noted anthocodial sclerites that were light orange, dark pink, or both; however, our materials are only orange. The coenenchymal sclerites are a bit smaller (up to 0.11×0.03 mm; Table 2) than previously described (up to 0.14-0.15×0.05 mm) (Verrill 1868: 398, Breedy and Guzmán 2007: 34). In addition, the Ecuadorian material has scattered small crosses not reported in this species before.

Leptogorgia flexilis (Verrill, 1868) (Figs 6, 7)

Synonymy. See Breedy and Guzmán (2007: 40). Newly collected examined material: MECN (Ant0034), Los Ahor- cados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 27 Feb. 2010, one whole colony. BEIM (0085), Los Ahorcados, Mana- bí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 27 Feb. 2010, one whole colony. MNCN (2.04/1171), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 5 m depth, 27 Feb. 2010, one whole colony. Additional examined materials: Litigorgia flexilis (Eugorgia flexi lis) MCZ 722 (4123), type material, Las Perlas (Panama), 6 to 8 m depth, 1860’s. Litigorgia flexilis NHM 69.4.15.15 (1946.1.14.74), type material, Toboguilla (Panama), 5 m depth, 1915. Leptogorgia flexilis YPM 569, Panama, no depth given, 1867-1868.

Description. The colonies measure up to 200 mm in length by 40 mm in width. The branching pattern is irregularly pinnate, mostly on a plane; branches are lank and bushy with long, slender, and flexible branches, drooping slightly at the ends (Fig. 6A). The unbranched distal twigs can reach up to 50 mm in length and 18 mm in diameter (Fig. 6A, B). The holdfast is circular, up to Fig. 5. – Leptogorgia diffusa (MNCN 2.04/1172) SEM photographs. 10 mm in diameter. The polyps retract within nearly Coenenchymal sclerites, A, spindles; B, captans; C, anthocodial flat polyp-mounds, closely distributed all around the sclerites, flattened rods. branches, with oblong apertures (Fig. 6B). The colour

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Fig. 7. – Leptogorgia flexilis (MNCN 2.04/1171) SEM photographs. Coenenchymal sclerites, A, spindles; B, captans; C, six-radiate; D, four-radiate.

sclerite type, up to 0.07 mm in length and 0.03 mm in width (Figs 6C, 7B). The spindles reach up to 0.1 mm in length and 0.03 mm in width, with 4-6 whorls of tubercles; they are straight or bent, some with a marked waist (Fig. 7A). Some crosses or four-radiates (up to 0.03×0.03 mm) (Fig. 7D) and six-radiates (up to 0.03×0.02 mm) are also found. The anthocodial sclerites are pink flattened rods up to 0.16 mm in length Fig. 6. – Leptogorgia flexilis (MNCN 2.04/1171). A, colony; B, detail of a branch; C, light micrograph of coenenchymal sclerites; and 0.08 mm in width, with short lobe-like marginal D, light micrograph of anthocodial sclerites. projections (Fig. 6D). of the colony is brown (Fig. 6A, B). The coenenchymal Geographic and bathymetric distribution. This sclerites are red, pink and yellow; some of them are species has been previously reported in California, El bicoloured (Fig. 6C). The capstans are the dominant Salvador and Panama at 5-30 m depth (Verrill 1868,

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Breedy and Guzmán 2007) (Fig. 25C), and thus our specimens represent a sizeable expansion to the south.

Remarks. Under water, the species is characterized by its decumbent or drooping branching pattern and its brown colonies. However, in small colonies the drooping habit is not as evident, and sometimes the holdfast and basal branches are an intense yellow. Crosses or four-radiate sclerites were not mentioned by Breedy and Guzmán (2007) in their revision of different type materials (e.g. deposited at MZC and YPM). However, they are evident in our collections and in the type specimens at NHM. This type of sclerite had been noted by Verrill (1868) as well, and it should be considered diagnostic for the species.

Leptogorgia obscura Bielschowsky, 1929 (Figs 8, 9)

Synonymy. See Breedy and Guzmán (2007: 57). Newly collected examined material: MECN (Ant0011), Los Ahor- cados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 27 Feb. 2010, three whole colonies. MECN (Ant0028), Los Frailes, Manabí (Ecuador), 1°30’14”S 80°48’33”W, 10 m depth, 17 Feb. 2012, one whole colony. BEIM (CRO-0068), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 27 Feb. 2010, one whole colony. BEIM (CRO-0069), Isla de Salango, Manabí (Ecuador), 1°35’55.13”S 80°52’0.01”W, 7 m depth, 19 Nov. 2011, one whole colony. MNCN (2.04/486), Isla de Salango, Manabí (Ecuador), 1°35’55.13”S 80°52’0.01”W, 7 m depth, 19 Nov. 2011, one whole colony. MNCN (2.04/487), Los Frailes, Manabí (Ecuador), 1°30’14”S 80°48’33”W, 10 m depth, 17 Feb. 2012, one whole colony. UTI (MZUTI-Inv03), Isla de Salango, Manabí (Ecuador), 1°35’55.13”S 80°52’0.01”W, 7 m depth, 19 Nov. 2011, one whole colony. UTI (MZUTI-Inv05), Isla de Salan- go, Manabí (Ecuador), 1°35’55.13”S 80°52’0.01”W, 13 m depth, 17 Feb. 2012, one whole colony. MZB (2016-2992), Isla de Salango, Manabí (Ecuador), 1°35’55.13”S 80°52’0.01”W, 13 m depth, 19 Feb. 2012, one whole colony. MZB (2016-2993), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 27 Feb. 2010, one whole colony. MECN (Ant0043), Los Frailes, Manabí (Ecuador), 1°30’14”S 80°48’33”W, 15 m depth, 23 May. 2013, one whole colony. MECN (Ant0044), Islote La Viuda, Manabí (Ecua- dor), 1°26’5.95”S 80°46’7.30”W, 15 m depth, 23 Nov. 2013, one whole colony. MECN (Ant0049), Punta Machalilla, Manabí (Ec- uador), 1°28’33.53”S 80°47’38.04”W, 13 m depth, 24 Nov. 2013, one whole colony.

Description. The colonies are up to 164 mm in length by 75 mm in width. The branching pattern is irregularly dichotomous; branches are bushy, closely ramified and rigid (Fig. 8A). The unbranched distal twigs can reach up to 5 mm in length and 2.3 mm in diameter, are compressed proximally, being more cylindrical and slightly tapered at the ends (Fig. 8A, B). The holdfast is circular, up to 14 mm in diameter. The polyps retract within prominent polyp-mounds leaving oblong apertures, and are closely distributed all around the branches (Fig. 8B). The colour of the colony is purple (Fig. 8A, B). The coenenchymal scle- Fig. 8. – Leptogorgia obscura (MECN Ant0011). A, colony; B, de- rites are red (Fig. 8C, D). The spindles are the domi- tail of a branch; C, light micrograph of coenenchymal sclerites; D, nant sclerite type, up to 0.13 mm in length and 0.04 light micrograph of anthocodial sclerites. mm in width, with 4-5 whorls of tubercles; they are straight or bent, some with a marked waist (Figs 8C, sclerites are orange flattened rods, up to 0.07 mm in 9A). The capstans reach up to 0.08 mm in length and length and 0.02 mm in width, with lobe-like marginal 0.04 mm in width (Figs 8C, 9B). Some crosses up to projections (Figs 8D, 9D). 0.06 by 0.04 mm are found (Fig. 9C). The anthocodial

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Fig. 10. – Pacifigorgia cribrum (MNCN 2.04/1170). A, colony; B, detail of a branch; C, light micrograph of sclerites.

Remarks. According to Bielschowsky (1929) and Fig. 9. – Leptogorgia obscura (MECN Ant0011) SEM photographs. Breedy and Guzmán (2007: 57), capstans are the Coenenchymal sclerites, A, spindles; B, captans; C, four-radiates; dominant sclerites in the type material. However, spin- D, anthocodial sclerites, flattened rods. dles are the most common sclerites in our specimens. Crosses or four-radiates also occur, a type of sclerite Geographic and bathymetric distribution. Lepto not reported in this species before. gorgia obscura was known previously only from the type locality (Bahía de Caráquez, Ecuador) (Biels- Genus Pacifigorgia Bayer, 1951 chowsky 1929; Fig. 25D). There is an unpublished record from Baja California (Harden 1979), although Synonymy. See Breedy and Guzmán (2002: 791). Breedy and Guzmán (2007) note that this record should be verified. The only reliable previously known depth Pacifigorgia cribrum (Valenciennes, 1846) range for the species was 4-5 m (Bielschowsky 1929), (Figs 10, 11) which we now expand to 15 m. This species is quite abundant in the study area on Synonymy. See Breedy and Guzmán (2002: 804). rocky bottom areas and usually grows along with Lep Newly collected examined material: MECN (Ant0035), Los Ahor- togorgia alba. cados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 15 m depth,

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- 35 yes 0.17×0.05 0.08×0.04 0.06×0.05 0.11×0.02 one or more slightly raised squarish/ 2×0.9 pink and yellow Pacifigorgia irene reddish with yellow red, lemon yellow and orange

2 not flat orange 0.16×0.04 0.12×0.04 0.09×0.04 0.09×0.02 light yellow one or more 0.08 by 0.06 brown or yellow oblong/ 55×6, 10×5 Pacifigorgia stenobrochis

- collected in this study, on the coast of Ecuador. no yes 1-3 two 9×4 orange 0.14×0.04 0.09×0.04 0.14×0.02 prominent pink and light yellow yellow with light brown Pacifigorgia flavimaculata Pacifigorgia rounded-square, oblong 19×5,

Fig. 11. – Pacifigorgia cribrum (MNCN 2.04/1170). SEM photographs. Coenenchymal sclerites, A, spindles; B, captans; C, an- 25 no yes thocodial sclerites, flattened rods. two 0.13×0.04 0.08×0.03 0.07×0.04 0.10×0.02 light yellow slightly raised 27 Feb. 2010, one whole colony. MECN (Ant0036), Los Ahorca- red and light yellow square / 2×2.8, 2.5×3 Pacifigorgia cribrum dos, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 20 m depth, 28 reddish (with yellow) Feb 2010, one whole colony. MECN (Ant0037), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 15 m depth, 27 Feb. 2010, one whole colony. BEIM (0088), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 25 m depth, 28 Feb. 2010, one whole colony. BEIM (0089), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 20 m depth, 28 Feb. 2010, one whole

colony. MECN (Ant0046), El Burbullón, Manabí (Ecuador), 1°28’23.59”S 80°51’23.04”W, 25 m depth, 23 Nov. 2013, one no whole colony. 6-9 yes several yellow 0.1×0.03 0.1×0.04 0.07×0.03 0.12×0.02 Comparative general features of the species genus

Additional examined materials: Rhipidigorgia cribrum MNHN-IK slightly raised

1661, type material, (New Zealand), no depth given, 1839. Pacifig brownish-orange

orgia cribrum MZC 261 (4014), Cape St. Lucas, Baja California, Pacifigorgia rubicunda (Mexico), no depth given, 1859-1861. MCZ 36264, Marcial Point square or oblong/ 10×3, 5.5×3 pink, orange and lemon yellow

(Mexico), no depth given, data unknown. Pacifigorgia cribrum Table 3. – USNM 49382 (Costa Rica), no depth given, Mar. 1927, F.M. Bayer (Id.). Gorgonia cribrum NMH 58.5.15.237 (Australia), no depth given, data unknown.

Description. The colonies are up to 50 mm in length –2 by 73 mm in width, and are formed by two fans: the first fan is the largest and the second one radiates from the holdfast and extends in parallel together with the first fan, until a certain point where both may fused (Fig. 10A). The holdfast is very small, up to 4 mm in diameter. The branches are squarish, ranging from 0.6 Characters Colour of colony Number of fans Number of meshes cm Mesh shape and maximum mesh size (mm) Polyp-mounds Colour of sclerites Bicolour sclerites Acute spindles max. size (mm) Blunt spindles max. size (mm) Capstan max. size (mm) Crosses Size and colour of anthocodial (mm)

SCI. MAR. 80(3), September 2016, 000-000. ISSN-L 0214-8358 doi: http://dx.doi.org/10.3989/scimar.04392.14A 12 • M.M. Soler-Hurtado et al. to 0.8 mm in diameter (25 meshes cm–2), and the end- branchlets are short, up to 2 mm long. The network is fine and regular; it is formed mostly by square meshes (2×2.8 mm by 2.5×3 mm) (Fig. 10B). There are no midribs; nevertheless, adult specimens have large, slightly compressed principal branches, which arise from near the base, and diverge through the fan, but often for no more than a quarter of the height. The polyps retract within slightly raised polyp-mounds with asterisk- like apertures, placed in multiple rows all around the branches. The colour of the colony is reddish intermingled with yellow. The coenenchymal sclerites are red, light yellow, and bicoloured (Fig. 10C). They are spindles (0.13×0.04 mm) having acute ends and 4-6 whorls of tubercles; blunt spindles (0.08×0.03 mm) with 4 whorls of tubercles (Figs 10C, 11A); and capstans (0.07×0.04 mm) with tuberculate ends (Figs 10C, 11B). The dominant sclerite types are acute, straight or bent spindles, some with a marked waist. The anthocodial sclerites are light yellow flattened rods, up to 0.10 mm in length and 0.02 mm in width, with smooth or slightly lobed borders (Figs 10C, 11C).

Geographic and bathymetric distribution. Pacifi gorgia cribrum has been previously reported in Mexico (Breedy and Guzmán 2002) (Fig. 25E), with additional questionable records from New Zealand (MNHN-IK 1661), Australia (NMH 58.5.15.237), and Costa Rica (e.g. USNM 49382, collected in 1927 and identified by M.F. Bayer). In this study we observed this species at 15-30 m depth. A depth range was lacking for it in the literature.

Remarks. According to Bayer and Macintyre (2001) and Breedy and Guzmán (2002), Pacifigorgia cribrum, P. adamsii, P. agassizii and P. rutila may represent a group sharing a set of morphological features, consist- ing of fine, regular and closely anastomosed networks. In addition, Pacifigorgia cribrum is morphologically close to P. arenata, both species having a similar colony colour and sclerome characteristics (red, yellow, bicoloured coenenchymal sclerites, and yellow anthocodial sclerites up to 0.10-0.13 mm in length; Table 3). However, the two species differ in mesh features (presence of midrib and higher meshes in P. arenata, see Breedy and Guzmán 2002). In fact, one of the principal problems in recording the possible morphological and chromatic variability of P. arenata is that the only known specimen is the holotype, which is deposited in Paris (MNHN), and there are some fragments in the Smithsonian (USNM 49567).

Pacifigorgia flavimaculata Breedy and Guzmán, 2003 (Figs 12, 13)

Newly collected examined material: MECN (Ant0021), Sali- Fig. 12. – Pacifigorgia flavimaculata (MNCN 2.04/1178). A, colo- nas, Santa Elena (Ecuador), 2°12’50.01”S 80°56’5.93”W, 15 m ny; B, detail of a branch; C, light micrograph of sclerites. depth, 10 Dec. 2011, one whole colony. MECN (Ant0022), Sali- nas, Santa Elena (Ecuador), 2°12’50.01”S 80°56’5.93”W, 18 m depth, 11 Dec. 2011, one whole colony. BEIM (0075), Salinas, 2010, one whole colony. MNCN (2.04/490), Los Frailes, Manabí Santa Elena (Ecuador), 2°12’50.01”S 80°56’5.93”W, 17 m depth, (Ecuador), 1°30’14”S 80°48’33”W, 10 m depth, 13 March 2012, 10 Dec. 2011, one whole colony. BEIM (0070), Los Ahorcados, one whole colony. MNCN (2.04/491), Salinas, Santa Elena Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 27 Feb. (Ecuador), 2°12’50.01”S 80°56’5.93”W, 17 m depth, 10 Dec.

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when alive and yellow intermingled with light brown and light purple when dry. The coenenchymal sclerites are pink and light yellow, some of them bicoloured (Fig. 12C). They are spindles (0.14×0.04 mm) having acute ends and 4-5 whorls of tubercles; blunt spindles (0.11×0.04 mm) with 4 whorls of tubercles (Figs 12C, 13A, B), and capstans (0.09×0.04 mm) with tuberculate ends, warty or smooth (Figs 12C, 13B). The dominant sclerite types are straight or bent spindles, some of them with a marked waist. The anthocodial sclerites are orange flattened rods, up to 0.13-0.14 mm in length and 0.02 mm in width, with pointed projections, and acute and warty ends (Figs 12C, 13C).

Geographic and bathymetric distribution. Pacifi gorgia flavimaculata has only been reported from the type locality in Costa Rica at 3 m depth (Breedy and Guzmán 2003) (Fig. 25F). In the present study, it was observed at a 3-20 m depth. Breedy and Guzmán (2003) pointed out that this species was only observed in the type locality (Punta Salsipuedes, Costa Rica), despite a significant sampling effort in the neighbouring areas. Therefore, our record from Ecuador is important in defining the geographic (and bathymetric) distribution of this species.

Remarks. Colonies of this species mainly form loose and irregular networks, sometimes almost pseudoanas- tomosed, yellow or brown in colour but with purplish or yellowish spots around a prominent polyp-mound, making the species easily recognizable. The Ecuadorian specimens have a quite constant set of features in comparison with the type specimens Fig. 13 – Pacifigorgia flavimaculata (MNCN 2.04/1178) SEM pho- examined (Breedy and Guzmán 2003, pers. observ.), tographs. Coenenchymal sclerites, A, acute spindles; B, capstans; C, anthocodial sclerites, flattened rods. except for the size of the anthocodial sclerites, which are longer (up to 0.13-0.14 mm, instead of up to 0.08- 2011, one whole colony. MZB (2016-2994), Los Frailes, Manabí (Ecuador), 1°30’14”S 80°48’33”W, 10 m depth, 20 Feb. 2012, 0.09 mm) (see Breedy and Guzmán 2003: 23, Table 3). one whole colony. UTI (MZUTI-Inv07), Los Frailes, Manabí (Ecuador), 1°30’14”S 80°48’33”W, 10 m depth, 20 Feb. 2012, Pacifigorgia irene Bayer, 1951 one whole colony. UTI (MZUTI-Inv08), Los Frailes, Manabí (Ec- (Figs 14, 15) uador), 1°30’14”S 80°48’33”W, 15 m depth, 10 March 2012, one whole colony. MECN (Ant0047), El Chichó, Manabí (Ecuador), 1°31’15.39”S 80°49’21.37”W, 20 m depth, 24 Nov. 2013, one Synonymy. See Breedy and Guzmán (2002: 825). whole colony. MECN (Ant0048), Punta Machalilla, Manabí (Ec- uador), 1°28’33.53”S 80°47’38.04”W, 13 m depth, 24 Nov. 2013, Newly collected examined material: BEIM (0091), Los Ahorca- one whole colony. dos, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 8 Dic. 2011, one whole colony. MNCN (2.04/1174), Punta Mala, Additional examined materials: Pacifigorgia flavimaculata MCZ Manabí (Ecuador), 1°33’41.37”S 80°50’8.79”W, 12 m depth, 16 51922, type material, Punta Salsipuedes (Costa Rica), 3 m depth, Feb. 2012, one whole colony. MZB (2016-2995), Los Ahorcados, 22 Jan 1994. Manabí (Ecuador), 1°40’44”S 80°50’08”W, 15 m depth, 15 April 2012, one whole colony. MECN (Ant0062), Los Frailes, Manabí (Ecuador), 1°30’14”S 80°48’33”W, 15 m depth, 10 March 2012, one whole colony. MECN (Ant0054), El Chichó, Manabí (Ecua- Description. The colonies reach up to 105 mm in dor), 1°31’15.39”S 80°49’21.37”W, 20 m depth, 24 Nov. 2013, one length by 70 mm in width and are formed by two par- whole colony. MECN (Ant0055), Islote La Viuda, Manabí (Ecua- allel fans. The holdfast was not observed (Fig. 12A). dor), 1°26’5.95”S 80°46’7.30”W, 15 m depth, 23 Nov. 2013, one The branches are cylindrical, ranging from 1.5-2.2 whole colony. mm in diameter (1-3 meshes cm–2). The branches arise Additional examined materials: Pacifigorgia irene USNM 49379, directly from the base and have incomplete anastomo- type material, Nicoya (Costa Rica), no depth given, Mar 1927; ses that form a loose, open and irregular network. The USNM 33611, type material, Costa Rica, no depth given, data unknown. Pacifigorgia irene MZC 36232, Gulf of Fonseca (Costa meshes are rounded-square, oblong or triangular (19×5 Rica), no depth given, data unknown. mm, 9×4 mm) (Fig. 12B). There are no distinct midribs. There are long end-branchlets (up to 11 mm). The Description. The colonies reach up to 600 mm in polyps retract within prominent polyp-mounds having length by 500 mm in width, and are formed by one rounded apertures, placed all around the branches in to several fans (Fig. 14A). The holdfast was not ob- multiple rows. The colour of the colony is light brown served. The branches are slender, ranging from 0.3

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length and 0.02 mm in width, with smooth or slightly lobed borders (Fig. 14C).

Geographic and bathymetric distribution. Pacifi gorgia irene has been previously reported in Panama and Costa Rica at 12-33 m depth (Bayer 1951, Breedy and Guzmán 2002, 2003) (Fig. 25G). Our finding represents a considerable southerly extension of its known distribution.

Remarks. This species is easily recognized by its characteristic morphology, showing a wide fan or fans, marked and thick midribs and small meshes. Some coenenchymal sclerites have been considered here as elongated capstans instead of blunt spindles, as com- monly described. The morphology of some sclerites are not always easy to define in this taxon, and they can also be considered transitional forms of blunt spindles, depending on the development of the two opposite distal processes on the longitudinal axis with respect to

Fig. 14. – Pacifigorgia irene (MNCN 2.04/1174). A, colony; B, detail of a branch; C, light micrograph of sclerites. to 0.7 mm in diameter (about 35 meshes cm–2). The branches form a fine and regular network. The network consists of small and squarish meshes (usually up to 1.2 by 0.9 mm) (Fig. 14B). The fans have several stout, rounded midribs. There are short end-branchlets (<1 mm long). The polyps retract within slightly raised and crowded polyp-mounds, placed all around the branches. The colour of the colony is reddish intermingled with yellow both when alive and dry, with a slight discoloration at the edges of the colony. The coenenchymal sclerites are red, lemon yellow and orange, some of them bicoloured (Fig. 14C). They consist of long spindles (0.17×0.05 mm) having acute ends and 5-6 whorls of tubercles (Figs 14A, 15A), and capstans (0.08×0.04 mm) with tuberculated ends (Fig. 15 B). In our specimens the dominant sclerite type are acute straight or bent spindles, some with a marked waist. Some crosses up to 0.06 by 0.05 mm occur as well (Fig. 15C). The anthocodial sclerites are light Fig. 15. – Pacifigorgia irene (MNCN 2.04/1174) SEM photographs. yellow and light pink flattened rods, up to 0.11 mm in Coenenchymal sclerites, A, spindles; B, captans; C, four-radiates.

SCI. MAR. 80(3), September 2016, 000-000. ISSN-L 0214-8358 doi: http://dx.doi.org/10.3989/scimar.04392.14A Gorgoniidae from Ecuador • 15 the two central whorls with alternate tubercles (Vargas et al. 2010a). In any case, if these sclerites are considered blunt spindles, they would have equivalent forms with respect to the original description of this species. In situ specimens seen during this study show a more intense reddish and yellow colour than Costa Rican specimens. Bayer (1951) described the colour of the colony as dark purple with greenish borders, but this is not the case in the Ecuadorian material, where colour only fades slightly at the edges of the fans.

Pacifigorgia machalilla n. sp. (Figs 16-18)

Examined material: Holotype: MECN (Ant0061), Cope, Manabí (Ecuador), 1°43’34”S 80°59’85”W, 23 m depth, 8 Dic. 2012, one whole colony without holdfast. Paratypes: MBZ (2016-2996), Los Ahorcados, Manabí (Ecuador), 1°40’44”N 80°50’08”W, 10 m depth, 1 Dic. 2011, one whole colony. MNCN (2.04/1181), Cope, Manabí (Ecuador), 1°43’34”S 80°59’85”W, 23 m depth, 8 Dic. 2012, a fragment of the colony. Other material: UTI (Inv262), Cope, Manabí (Ecuador), 1°43’34”S 80°59’85”W, 23 m depth, 8 Dic. 2012, one whole colony. BEIM (0095), Isla de la Plata, Manabí (Ecuador), 1°16’25.84”S 81° 4’11.70”W, 22 m depth, 22 Feb. 2012, two whole colonies. MECN (Ant0058), Los Ahorcados, Manabí (Ecuador), 1°40’44”N 80°50’08”W, 10 m depth, 27 Feb. 2010, one whole colony. MECN (Ant0059), Los Ahorcados, Manabí (Ecua- dor), 1°40’44”N 80°50’08”W, 10 m depth, 1 Dic. 2011, one whole colony.

Description of the holotype. The colony is formed by a single fan 250 mm in length by 290 mm in width (Fig. 16A). The holdfast is circular, up to 15 mm in diameter. The branches are cylindrical, ranging from 1.5-2 mm in diameter (7-9 meshes cm–2) and the end-branchlets reach up to 9 mm in length, and have blunt tips. The network is regular and complete, and is Fig. 16. – Pacifigorgia machalilla n. sp. Holotype MECN (Ant0053). formed of mostly square and rectangular meshes (4×5 A, colony; B, detail of anastomosed branches; C, light micrograph mm, 10×3 mm); in some cases, meshes are oblong (Fig. of sclerites; D, light micrograph of anthocodial sclerites. 16B). There are five or six prominent, long and strong midribs, which divide into others that progressively ing parallel to the primary fan. In addition, in some fuse among the anastomosed structure of the mesh small colonies (about 100 mm in length) the midrib is (Fig. 16A). The polyps retract within slightly raised not very obvious or even absent, or is only formed by or flat polyp-mounds with slit-like apertures, placed in two short basal branches. multiple rows all around the branches. The colour of the colony is intense red-brown when dry and bluish- Geographic and bathymetric distribution. Pacifi grey when alive. The coenenchymal sclerites are pink, gorgia machalilla is only known from the type locality light yellow and orange, some of them bicoloured (Fig. in Cope, Los Ahorcados, and Isla de la Plata (conti- 16C). They are spindles (up to 0.20×0.04 mm) hav- nental coast of Ecuador), living on rocky bottoms in ing acute ends and 5-6 whorls of tubercles (Figs 16A, shallow waters at a depth of 10-23 m. 17A); blunt spindles are absent, and there are capstans (up to 0.7×0.03 mm) with tuberculate ends (Figs 16C, Etymology. The new species is dedicated to the 17B). The dominant sclerites are spindles, straight or National Park of Machalilla (Ecuador) and its staff bent, some of them with a distinct waist. There are for their constant support and help during field work. irregular crosses with different branch lengths (up to Name considered as a noun in apposition. 0.11×0.05 mm) (Fig. 17C). The anthocodial sclerites are pink, light orange and light yellow in colour, most Comparison with other Pacifigorgia species. Paci of them bicoloured; flattened rods (0.11×0.02 mm) figorgia machalilla is morphologically close to P. exi with short pointed or lobe-like marginal projections lis and P. firma, having similar networks with the pres- and acute or rounded ends also occur. There are also ence of a midrib, and similar mesh size (9-11.5 meshes platelets (0.06×0.02 mm) (Figs 16C, 17D). cm–2) and anastomosis. The network in P. machalilla is formed by almost square or rectangular meshes, Variability. This new species is fairly constant with 10×3 mm, 4×5 mm, and 12×3 mm, 6×2 mm in P. firma regard to colonial and sclerome characters. However, in (Table 4). Pacifigorgia exilis also has a similar type some specimens there are small secondary fans grow- of network (open or closed and regular); however, it

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Table 4. – Comparative morphological features of Pacifigorgia machalilla n. sp. with its closest congeners. Characters Pacifigorgia machalilla Pacifigorgia exilis Pacifigorgia firma Colour of colony intense red-brown red-orange light brown/ dark purple Number of fans one or two small several one (or two) Number of meshes cm–2 7-9 7 - 9 11.5 Mesh shape and maximum square -rectangular/ regular square - oblong / irregular rectangular / Mesh size (mm) 10×3 - 4×5 3×0.5 regular 12×3 - 6×2 Presence of midrib present present present Polyp-mounds slightly raised or flat slightly raised slightly raised Colour of sclerites pink /yellow/orange red / yellow red/pale yellow/orange Acute spindles max. size (mm) 0.20 length 0.04 width 0.11 length 0.05 width 0.11 length 0.05 width Blunt spindles max. size (mm) absent 0.09 length 0.04 width absent Capstan max. size (mm) 0.07 length 0.03 width 0.07 length 0.04 width 0.09 length 0.05 width Crosses or radiates present absent absent Anthocodial max. size (mm) 0.11 length 0.02 width 0.12 length 0.02 width 0.10 length 0.03 width Shape and colour of anthocodial flattened rods; pink/ orange / light flattened rods; yellow present; flattened rods; ligth orange rods; Bicolour sclerites yellow present; coenenchyme coenenchyme present; coenenchyme and anthocodial References This paper Breedy and Guzmán 2002 Breedy and Guzmán 2003 has smaller square or oblong meshes (0.5×3 mm). Ad- firma (up to 0.11×0.05 mm). Finally, P. machalilla has ditionally, P. machalilla has blunt spindles while they crosses or radiate forms, which are lacking in the other are absent in both P. exilis and P. firma, and the acute two species. spindles are larger in P. machalilla (up to 0.20×0.04 The molecular distance matrix (COII + Igr + COI mm) than in P. exilis (up to 0.11×0.05 mm) and P. + MutS) shows that, within the genus Pacifigorgia,

Fig. 17. – Pacifigorgia machalilla n. sp. Holotype MECN (Ant0053). SEM photographs. Coenenchymal sclerites, A, acute spindles; B, captans; C, irregular crosses or four-radiates and butterfly; D, anthocodial sclerites, flattened rods.

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Fig. 18. – Comparison of the anastomosed branches in Pacifigorgia machalilla n. sp., holotype (A) and Pacifigorgia irene (B).

Table 5. – Genetic divergence matrix for the species examined based on the COII + Igr + COI and MutS-1 gene sequence. 1 2 3 4 5 6 7 9 9 10 11 12 13 14 P. firma 1 P. rubicunda (1) 2 0.0127 P. rubicunda (2) 3 0.0128 0.0000 P. smithsoniana 4 0.0024 0.0114 0.0114 P. stenobrochis 5 0.0055 0.0108 0.0108 0.0042 P. catedralensis (1) 6 0.0006 0.0115 0.0115 0.0024 0.0042 P. catedralensis (2) 7 0.0006 0.0116 0.0116 0.0024 0.0043 0.0000 P. sculpta 8 0.0060 0.0114 0.0114 0.0048 0.0018 0.0048 0.0049 P. exilis 9 0.0133 0.0006 0.0006 0.0119 0.0118 0.0121 0.0122 0.0116 P. machalilla n. sp. (1) 10 0.0030 0.0108 0.0108 0.0006 0.0036 0.0018 0.0018 0.0039 0.0110 P. machalilla n. sp. (2) 11 0.0030 0.0108 0.0108 0.0006 0.0036 0.0018 0.0018 0.0039 0.0110 0.0000 P. machalilla n. sp. (3) 12 0.0030 0.0108 0.0108 0.0006 0.0036 0.0018 0.0018 0.0039 0.0110 0.0000 0.0000 P. bayeri 13 0.0012 0.0114 0.0114 0.0024 0.0042 0.0000 0.0000 0.0048 0.0120 0.0018 0.0018 0.0018 P. media 14 0.0139 0.0006 0.0006 0.0125 0.0112 0.0126 0.0128 0.0118 0.0000 0.0118 0.0118 0.0118 0.0125 P. irene 15 0.0036 0.0114 0.0108 0.0012 0.0042 0.0018 0.0018 0.0048 0.0120 0.0006 0.0006 0.0006 0.0024 0.0125 the genetic divergence among species was close to 1% there is a mutation in the first base of one triplet which or lower in most cases. The new species showed no changes Lys to Val, in COII for P. machalilla and P. intraspecific variation (Table 5). Despite the fact that irene. It may be interesting to extend the sequencing P. machalilla is closely related morphologically to P. based on COII in the search of additional variable seg- firma and P. exilis, as stated previously, the three spe- ments in the mtDNA of this group of species. cies are clearly separated molecularly. The maximum Despite its molecular similarities, there are clear genetic divergence is 0.3% between P. machalilla and morphological differences among these three species. P. firma, and is 1% between P. machalilla and P. exilis Pacifigorgia machalilla differs from P. smithsoniana (Table 5). However, Pacifigorgia machalilla is closely in having a midrib, and larger acute spindles (0.20×0.04 related molecularly to P. irene and P. smithsoniana vs 0.14×0.05 mm). Pacifigorgia machalilla differs (Breedy and Guzmán, 2004). The maximum genetic from P. irene in that its anastomosis forms a network divergence between P. machalilla and P. smithsoniana of almost square or rectangular meshes 10×3 mm, 4×5 or P. irene is 0.06%. However, in a thorough analysis mm (only 2×0.9 mm in P. irene), its branches are larger, of the sequences for these three species, we observed ranging from 1.5-2 mm in diameter (7-9 meshes cm–2), the following differences: 1) there is a silent mutation while the branches in P. irene are 0.5-0.7 mm in diam- in COI for P. machalilla and P. smithsoniana; and 2) eter (35 meshes cm–2), with longer end-branchlets (up

SCI. MAR. 80(3), September 2016, 000-000. ISSN-L 0214-8358 doi: http://dx.doi.org/10.3989/scimar.04392.14A 18 • M.M. Soler-Hurtado et al. to 9 mm long), while these are very short in P. irene, less than 1 mm (Fig. 18). Finally, bicolour anthocodial sclerites are a unique feature for P. machalilla.

Pacifigorgia rubicunda Breedy and Guzmán, 2003 (Figs 19, 20)

Newly collected examined material: MECN (Ant0023), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 27 Feb. 2010, two whole colonies. MECN (Ant0025), Punta Mala, Manabí (Ecuador), 1°33’41.37”S 80°50’8.79”W, 15 m depth, 15 Feb. 2012, three whole colonies. BEIM (0083), Isla de la Plata, Manabí (Ecuador), 1°16’25.84”S 81° 4’11.70”W, 22 m depth, 19 Feb. 2012, one whole colony. BEIM (0082), Los Frailes, Manabí (Ecuador), 1°30’14”S 80°48’33”W, 10 m depth, 10 March 2012, one whole colony. BEIM (0081), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 5 m depth, 27 Feb. 2010, one whole colony. MNCN (2.04/488), Isla de la Plata, Manabí (Ecuador), 1°16’25.84”S 81° 4’11.70”W, 20 m depth, 19 Feb. 2012, one whole colony. MNCN (2.04/489), Punta Mala, Manabí (Ecuador), 1°33’41.37”S 80°50’8.79”W, 12 m depth, 27 Feb. 2012, one whole colony. UTI (MZUTI-Inv04) Los Ahorca- dos, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 27 Feb. 2010, one whole colony. MZB (2016- 2997), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 5 m depth, 27 Feb. 2010, one whole colony. MZB (2016-2998) Isla de la Plata, Manabí (Ecuador), 1°16’25.84”S 81° 4’11.70”W, 20 m depth, 19 Feb. 2012, one whole colony.

Additional examined materials: Pacifigorgia ru bicunda MCZ 51917, type material, Gulf of Chiriquí (Panama), 14 m, Dec. 2001.

Description. The colonies reach up to 40 mm in length by 160 mm in width, and are formed by several fans. The secondary fans sprout from the main fan at a right angle and spread perpendicularly to form new fans. Several fans may reunite, adhering together and produc- ing square arrangements like beehives. A short stem divides close to the holdfast (Fig. 19A), which is up to 12 mm in diameter. The branches are cylindrical, ranging from 0.8-1 mm in diameter (9-11 meshes cm–2). The end-branchlets are short. The network is closed and regular; it is formed of mostly square or oblong meshes (10×3 mm, 5.5×3 mm) (Fig. 19B). The polyps retract within slightly raised polyp-mounds with slit-like apertures, placed in multiple rows all around branches. The colour of the colony is brownish-orange when alive or preserved and a conspicuous burnt sienna colour speck- Fig. 19. – Pacifigorgia rubicunda (MZB 2016-2997). A, colony; B, led with yellow when dry. The coenenchymal sclerites detail of a branch; C, light micrograph of sclerites. are pink, orange and lemon yellow, bicoloured and multicoloured (Fig. 19C). They are spindles (0.1×0.03 The anthocodial sclerites are yellow flattened rods up mm) having acute ends and 10-12 whorls of tubercles to 0.12 mm in length and 0.02 mm in width, with short (Figs 19C, 20A); blunt spindles (0.1×0.04 mm) with lobe-like marginal projections, and acute and warty or 4-5 whorls of tubercles (Figs 19C, 20B); and capstans smooth ends (Figs 19C, 20C). (0.07×0.03 mm) with tuberculate ends (Figs 19C, 20C). The dominant sclerite types are blunt spindles straight Geographic and bathymetric distribution. Pacifi or bent, some with a marked waist. Crosses are lacking. gorgia rubicunda was originally described by Breedy

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and Guzmán (2003) from Costa Rica (Fig. 25I). The geographic range of the species is here extended to Ecuador (Manabí). Its bathymetric distribution ranges from 5 to 30 m depth (Breedy and Guzmán 2003).

Remarks. There are some morphological differences between the specimens of P. rubicunda found in Ecuador and the original description of this species by Breedy and Guzmán (2003). The colour of sclerites is very similar (orange and yellow); however, in Ecuado- rian material most of the sclerites are pink (bicoloured or multicoloured) and colourless sclerites are lacking. If compared with the type material (6-9 meshes cm–2, 5-1.2×2.5-0.9 mm) (Breedy and Guzmán 2003), in our colonies the network is less compact, usually more elongate and larger, and there are no crosses. Finally, the size of sclerites and anthocodial rods seem to be somewhat larger in the type material (spindles 0.09-0.12×0.03-0.04 mm, capstans 0.04- 0.09×0.02-0.04 mm; anthocodial rods 0.14×0.05 mm) than in the Ecuadorian material (spindles 0.1×0.03 mm, capstans 0.07×0.03 mm; anthocodial rods 0.12×0.02 mm). In most cases, P. rubicunda was found form- ing colonies similar to beehives, irregularly stretching across the rocks. The midrib extends from the holdfast in parallel to the substrate, but the unique point of un- ion is the holdfast, which is diagnostic for this species (Breedy and Guzmán 2003: 43). Newly collected material of P. rubicunda from Ecuador permits the description of variability of some characters. The species is reported herein for the first time since its original description in Costa Rica.

Pacifigorgia stenobrochis (Valenciennes, 1846) (Figs 21, 22)

Synonymy. See Breedy and Guzmán (2002: 833). Newly collected examined material: BEIM (0090), Salinas, Santa Elena (Ecuador), 2°12’50.01”S 80°56’5.93”W, 15m depth, 9 Dic. 2011, a fragment of colony. MNCN (2.04/1176), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 12 Dic. 2011, a fragment of colony. MNCN (2.04/1177), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 13 m depth, 7 Dic. 2012, a fragment of colony. MZB (2016-2999), Punta Mala, Manabí (Ecuador), 1°33’41.37”S 80°50’8.79”W, 15 m depth, 12 Dic. 2012, a fragment of colony. MNCN (2.04/1175), Salinas, Santa Elena (Ecuador), 2°12’50.01”S 80°56’5.93”W, 15m depth, 11 Dic. 2011, a fragment of colony. MECN (Ant0065), Los Ahorcados, Manabí (Ecuador), 1°40’44”S 80°50’08”W, 10 m depth, 15 Mar. 2012, a fragment of colony. MECN (Ant0064), Punta Mala, Manabí (Ec- uador), 1°33’41.37”S 80°50’8.79”W, 15 m depth, 15 Dic. 2011, a fragment of colony. MECN (Ant0063), Punta Mala, Manabí (Ec- uador), 1°33’41.37”S 80°50’8.79”W, 13 m depth, 6 April 2012, a fragment of colony. MECN (Ant0051), El Chichó, Manabí (Ecua- dor), 1°31’15.39”S 80°49’21.37”W, 20 m depth, 24 Nov. 2013, a fragment of colony. MECN (Ant0052), El Burbullón, Manabí (Ec- uador), 1°28’23.59”S 80°51’23.04”W, 25 m depth, 23 Nov. 2013, a fragment of colony. MECN (Ant0053), Islote La Viuda, Manabí (Ecuador), 1°26’5.95”S 80°46’7.30”W, 15 m depth, 23 Nov. 2013, a fragment of colony. Additional examined materials: Rhipidigorgia stenobrochis MNHN-IK 1719, type material, New Zealand, no depth given, 1839. Pacifigorgia stenobrochis var. engelmanni (Horn 1860) MCZ 4042, Acapulco (Mexico), no depth given, 1856-1860; Pacifigorgia stenobrochis MCZ 28753, Mexico, no depth given, data unknown; Fig. 20. – Pacifigorgia rubicunda SEM photographs. Coenenchy- Gorgonia stenobrochis NHM 1930.6.17.12 St. George (Mexico), mal sclerites, A, spindles; B, captans; C, anthocodial sclerites, flat- no depth given, data unknown. Pacifigorgia stenobrochis USNM tened rods. 49378, Gulf of Nicoya (Costa Rica), no depth given, March 1927.

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Fig. 21. – Pacifigorgia stenobrochis (MNCN 2.04/1175). A, colony; B, detail of a branch; C, light micrograph of sclerites.

Pacifigorgia stenobrochis USNM 8847, Baja California (Mexico), no depth given, 1911. Fig. 22. – Pacifigorgia stenobrochis (MNCN 2.04/1175). SEM photographs. Coenenchymal sclerites, A, spindles; B, captans; C, crosses; D, anthocodial sclerites, flattened rods. Description. The colonies are up to 280 mm in length by 270 mm in width and are formed by a single lar or oblong (55×6 mm, 10×5 mm) (Fig. 21B). There fan that may divide into smaller fans. The holdfast was are no distinct midribs. There are long end-branchlets not observed (Fig. 21A). The branches are rounded, (up to 26 mm). The polyps retract within flat polyp- ranging from 1.1-2.3 mm in diameter (2 meshes cm–2). mounds, placed throughout the branches, on both sides. The branches arise directly from the base and have an The colour of the colony is light brown or dark yellow open and irregular network. The meshes are rectangu- when alive and yellow intermingled with brown. The

SCI. MAR. 80(3), September 2016, 000-000. ISSN-L 0214-8358 doi: http://dx.doi.org/10.3989/scimar.04392.14A Gorgoniidae from Ecuador • 21 coenenchymal sclerites are light yellow (Fig. 21C). There are long acute spindles (0.16×0.04 mm), straight or bent, some with a marked waist, having 8 whorls of tubercles, and they are the dominant sclerite types. In addition, there are blunt spindles (0.12×0.04 mm) with 6 whorls of tubercles (Figs 21C, 22A), and capstans (0.09×0.04 mm) with tuberculate ends (Figs 21A, 22B). Some crosses up to 0.08 by 0.06 mm are also found (Fig. 22C). The anthocodial sclerites are light orange in colour, with flattened rods up to 0.09 mm in length and 0.02 mm in width, with smooth or slightly lobed borders (Figs 21A, 22D).

Geographic and bathymetric distribution. Pacifi gorgia stenobrochis has been reported in Mexico, El Salvador, Nicaragua, Costa Rica, Panama, Peru (Ver- rill 1868, Breedy and Guzmán 2003) and Ecuador (current study) at 3-30 m depth. In the original description, New Zealand is indicated as the type locality, which appears to be an error (see Valenciennes 1846, Breedy and Guzmán 2002, 2003) (Fig. 25J). Generally, colonies of this species occur both soli- tary and mixed with other species in the same environment. The Ecuador records nicely connect the previously known localities from Mexico and Peru.

Remarks. The chromatic variability of the colonies and sclerites of this species has been known for a long Fig. 23. – Eugorgia daniana (MNCN 2.04/1179). A, colony; B, time (Hickson 1928, Breedy and Guzmán 2002, 2003). detail of a branch; C, light micrograph of sclerites. In the Ecuadorian material studied here, all the coenenchymal sclerites are yellow. Pink and grey sclerites noted by other authors are lacking. The colour of Ecuadorian colonies varies between yellow and orange hues, while it is said to be reddish purple and brown in previous descriptions. The presence of thick and robust branches, shape of the mesh and absence of the midrib are diagnostic for P. stenobrochis, and readily separate it from similar species, such as P. firma.

Genus Eugorgia Verrill, 1868

Synonymy. See Breedy et al. (2009: 8).

Eugorgia daniana Verrill, 1868 (Figs 23, 24)

Synonymy. See Breedy et al. (2009: 17). Newly collected examined material: MECN (Ant0033), Salinas, Santa Elena (Ecuador), 2°12’50.01”S 80°56’5.93”W, 15 m depth, 9 Dic. 2011, one whole colony. MNCN (2.04/1173), Salinas, Santa Elena (Ecuador), 2°12’50.01”S 80°56’5.93”W, 15 m depth, 10 Dic. 2011, one whole colony. MECN (Ant0060), Punta Gruesa, Manabí (Ecuador), 1°33’38.15”S 80°50’5.28”W, 18 m depth, 20 Feb. 2013, one whole colony. MECN (Ant0056), El Chichó, Manabí (Ecua- dor), 1°31’15.39”S 80°49’21.37”W, 20 m depth, 24 Nov. 2013, one whole colony. MNCN (2.04/1179), El Burbullón, Manabí (Ecua- dor), 1°28’23.59”S 80°51’23.04”W, 25 m depth, 23 Nov. 2013, one whole colony. Additional examined materials: Eugorgia daniana MCZ 7080, type material, Gulf of Nicoya (Costa Rica), no depth given, May 1868; MCZ 723, type material, Las Perlas (Panama), no depth given, 1866-1867; NHM 69.4.15.55, type material, Panama, no depth giv- Fig. 24. – Eugorgia daniana (MNCN 2.04/1179) SEM photographs. en, data unknown. Eugorgia daniana MCZ 02138, Cali (Colombia), A, coenenchymal sclerites, spindles; B, double disc; C, crosses; D, disc-spindles; E, complete double disc.

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3 m depth, Dec. 1979; USNM 59084 Gulf of Guayaquil (Ecuador), 20 m depth, Sept. 1966. DISCUSSION Description. The colonies are up to 110 mm in length Seven of the species identified are new records for by 190 mm in width and arise from a circular holdfast, the octocoral fauna of Ecuador (Table 1, 2): Leptogor 0.5 mm in diameter. They are profusely and pinnately gia diffusa, L. flexilis, Pacifigorgia cribrum, P. flavi branched in multiple planes. Main stems are 3-5 mm maculata, P. irene, P. rubicunda and P. stenobrochis. in diameter, compressed, and very short (up to 10 mm Another three species, Leptogorgia alba, L. obscura long) or absent (Fig. 23A). The branches arise directly and Eugorgia daniana, had already been reported in from the base, and are cylindrical, ranging from 0.8 to Ecuador by other authors (Bielschowsky 1929, Breedy 1.2 mm in diameter. Some of the branches are pseudoa- and Guzmán 2007). The new species described here, nastomosed (anastomosis to the coenenchyme level, but Pacifigorgia machalilla, requires additional records not including the axes). The unbranched distal twigs can to better establish its geographical and bathymetric reach up to 15 mm in length and are blunt at the ends distribution. (Fig. 23A, B). In some branches, there are numerous and These species, together with Leptogorgia maria evident yellow, longitudinal grooves. The polyps retract rosae Soler-Hurtado and López-González, 2012, within prominent polyp-mounds, leaving a small bilabi- Eugorgia ahorcadensis and L. aequatorialis (Biels- ate aperture; they occur in multiple rows on all sides of chowsky, 1929), constitute the current list of gorgoniid the branches, sometimes having yellow spots. Colonies species from the continental coast of Ecuador. Despite are dark orange or bright red streaked with bright yellow the recent contribution of Breedy et al. (2013) suggest- on the branches or on the polyp-mounds (Fig. 23B). The ing the synonymization of E. ahorcadensis with E. coenenchymal sclerites are red, yellow or bicoloured nobilis based on the consideration of an enlargement of (Fig. 23C). They are double discs reaching up to 0.07 morphology and chromatic variability of the latter, re- mm in length, 0.06 mm in width (Figs 23C, 24B). The cent molecular analyses based on the study of both type capstans reach up to 0.08 mm in length, 0.05 mm in materials demonstrated that they should be considered width (Fig. 23C). The dominant sclerite types are spin- separate taxa (Soler-Hurtado et al. unpublished). In dles (up to 0.15 mm in length, 0.05 mm in width); disc- addition, to complete the species list for Ecuadorian spindles (up to 0.14 mm in length, 0.05 mm in width) waters, the species described above for the Galapagos are also abundant, with 4-6 whorls of warty tubercles; Islands, Pacifigorgia darwinii (Hickson, 1928), P. they are straight or bent, some with a marked waist (Fig. dampieri Williams and Breedy, 2004, P. symbiotica 24A, D). The crosses are up to 0.14 mm in length, 0.06 Williams and Breedy, 2004 and P. rubripunctata Wil- mm in width (Figs 23C, 24C). No anthocodial sclerites liams and Breedy, 2004, should be added. were obtained. According to Bayer (1961), gorgonian corals systematics relies on a combination of characters such as Geographic and bathymetric distribution. This spe- axial skeleton (if any), colonial form, polyp arrange- cies has a wide distribution along the eastern Pacific. ment and sclerite morphology. However, the often un- It has been recorded in USA, Mexico, El Salvador, wieldy older literature with no or neglected illustrations Nicaragua, Costa Rica, Panama, Colombia, Ecuador and lost or badly preserved type material have played a (Galapagos and the Gulf of Guayaquil) and Peru (Fig. major role in the taxonomic confusion around octocoral 25K), at 3-30 m depth (Verrill 1868, Breedy et al. systematics (Sánchez 2004, Soler-Hurtado and López- 2009, present record). González 2012). Currently, variation in sclerite size, The only specimen of Eugorgia daniana previously sculpture, and coloration and the relative proportion of found in continental Ecuadorian waters was collected the different sclerite types are the main criteria for spe- in the Gulf of Guayaquil (identified by Bayer in 1966, cies delimitation in Gorgoniidae (Williams and Lindo and published by Breedy et al. 2009). Our record from 1997, Breedy 2001, Sánchez et al. 2007, Vargas et al. Manabí and Santa Elena is the second finding of this 2010a). These characters vary widely and the ranges species in continental Ecuador, providing information of variation have not been established for all gorgoniid about the possible existence of widespread populations taxa, hindering a robust taxonomy for the group. of this species in this area. Currently, Eugorgia dani The description of sclerite type can sometimes be ana and E. ahorcadensis Soler-Hurtado and López- difficult, because of continuous variation in size and González, 2012 are the only species of this genus found ornamentation within and between species (Williams in Ecuadorian waters. and Lindo 1997, Sánchez et al. 2003, Vargas et al. 2010a). This continuum represents a major obstacle to Remarks. Differentiation between E. daniana and the assignment of size intervals or ranges that could be E. aurantica is difficult at first sight due to a similar used to define and separate possible species. Vargas et branching pattern and colour of their colonies. al. (2010b) proposed for Pacifigorgia the combined use However, E. aurantica has polyp-mounds with of continuous and discrete morphological characters, characteristic yellow rings and lacks disc-spindles. Eu in order to define the relationships between species at gorgia daniana, E. aurantica and E. multifida Verrill, different resolution level, in an integrative approach 1870 form the “daniana-group” (Breedy et al. 2009, with molecular data. 2013), which includes species with irregular pinnate Despite the rebirth of invertebrate systematics due to branching, prominent polyp-mounds and a similar col- the ever-increasing availability of DNA sequence char- our of colonies and sclerites. acters (Mallet and Willmott 2003, Sánchez 2004, Due-

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Fig. 25. – Geographical distribution of the Leptogorgia, Pacifigorgia and Eugorgia species mentioned in the paper. The solid circle shows the relative position of the species in the countries where they have been recorded. The empty circles indicate the first record for the species in Ecuador. ñas and Sánchez 2009), evaluation of the effectiveness (2011) showed that congeners of the genera Isidella, of molecular barcodes in octocorals is largely hindered Keratoisis and Lepidisis were identical at COI and Igr1 by lack of knowledge regarding species boundaries in sequences, and only varied 0%-2% for MutS. these organisms. One reason for this is that the rate of In this study, we found that pairs of morphologi- octocoral mitochondrial gene evolution is very slow. It cally different species, Pacifigorgia exilis and P. media is estimated to be 10-100× slower than in other metazo- (Verrill, 1864), and P. bayeri Breedy, 2001 and P. cat ans (Chen et al. 2009, Brockman and McFadden 2012), edralensis Breedy and Guzmán, 2004, showed no in- resulting in insufficient resolution to discriminate spe- terspecific variation (0%, in COII+Igr+COI+MutS, cf. cies within many genera (Sánchez et al. 2003, Lepard Table 4); while others, P. firma and P. catedralensis, 2003 in Cairns and Bayer 2005). Specimens identified P. rubicunda and P. exilis, and P. rubicunda and P. me as different morphospecies can share the same barcode, dia, showed a very low genetic divergence (0.006%). which should motivate additional taxonomic work to In fact, the maximum genetic divergence observed in test species boundaries and quantify intraspecific mor- this study is 1.3%, between P. media and P. firma, and phological and molecular variation (McFadden et al. between P. exilis and P. firma, confirming the reduced 2010). For instance, Sánchez et al. (2003), exploring variability in the genetic regions examined. mitochondrial genes ND2, ND6 and MutS, observed The analysis combining MutS and Igr1-COI for that in Eunicea spp. and Plexaura spp. the number DNA barcoding indicates that these markers are not of substitutions supporting nodes was low (less than always suitable and conclusive for species-level iden- 0.005 substitutions/site), and the clade formed by these tification of eastern Pacific octocorals (Vargas et al. two genera could be considered unresolved from a 2014). However, we can say that these regions can at molecular point of view. In addition, McFadden et al. least define different species groups. This may serve

SCI. MAR. 80(3), September 2016, 000-000. ISSN-L 0214-8358 doi: http://dx.doi.org/10.3989/scimar.04392.14A 24 • M.M. Soler-Hurtado et al. as a supporting tool for morphological findings among Universidad Tecnológica Indoamérica to JM. This species within and between groups. research received support from the SYNTHESYS According to our results, based on strong morpho- Project (http://www.synthesys.info/), financed by the logical data and molecular tests with sufficient resolu- European Community Research Infrastructure Action tion for this set of octocoral species, we propose Paci under the FP7 Capacities Programme, and “Ernst Mayr figorgia machalilla as a new species. Travel Grants In Animal Systematics” for visiting In conclusion, we recommend the use of molecu- MCZ at Harvard University. This research was partial- lar tools as a necessary complement to morphological ly supported by a grant from the Spanish Ministry of identification for future descriptions of new species. Economy and Competitiveness (CTM2014-57949-R). Although mitochondrial markers are known to evolve Our special thanks to Gonzalo Giribet and Adam J. at much lower rates than in other zoological groups, Baldinger (MCZ) and Aude Andouche (MNHN), they still provide information that should not be ne- Andrew Cabrinovic and Miranda Lowe (NHM), Eric glected. These mitochondrial markers can be comple- Lazo-Wasem and Lourdes Rojas (YPM), and Stephen mented with nuclear regions (e.g. 28S, ITS and SRP54, Cairns and Herman H. Wirshing (Smithsonian, MNH) among others) that have been demonstrated to be es- for helping in our museum work. We thank Santiago pecially useful for some octocoral families and genera Villamarín (MECN), Machalilla National Park and (Sánchez et al. 2007, McFadden et al. 2014, Wirshing Ministerio del Ambiente of Ecuador (Manabí) for and Baker 2015). collection permits; and Michel Guerrero and his team Although much more work is needed to fully under- (Exploramar Diving) for his special interest and collab- stand the morphological diversification of octocorals, a oration since we began our research. Special thanks to combination of molecular and morphological data is a Micaela Peña for her unconditional help, and to Alicia very promising approach to disentangling phylogenetic Maraver, Valeria García and Damián Ramírez for their relationships among species (Breedy et al. 2013, Mc- help on the collection expeditions, their friendship and Fadden and Van Ofwegen 2013, López-González et al. their hard work. Thanks also to Eva Andrés Marruedo 2015) and intraspecific population ecology (Calderón and Mónica Flores for their help during the subsequent et al. 2006, Prada et al. 2008, Prada and Hellber 2013). fieldwork. Finally, we thank one anonymous referee Molecular and morphological analytic tools will be es- for his/her suggestions for improving our manuscript. sential to quantify the variety of evolutionary pathways within these groups (Sánchez et al. 2003, McFadden et REFERENCES al. 2006, Concepción et al. 2010). However, even when combined, the two approach- Alderslade P. 1998. Revisionary systematics in the gorgonian family Isididae, with description of numerous new taxa (Coelen- es do not seem to fully solve the taxonomic problems terata, Octocorallia). Rec. West. Aust. Mus. 55: 1-359. in this group, and more informative characters in both Bayer F.M. 1951. 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